ObjectiveTo explore the light response, retinal inflammation and apoptosis of the retinal ganglion cells (RGCs) 1 year after the new type of channelrhodopsin PsCatCh2.0 was transfected into the retina of rd1 mice. MethodsTwenty-four male rd1 mice were randomly divided into rd1 experimental group and rd1 control group, 12 mice in each group. 1.5 μl of recombinant adeno-associated virus (rAAV)2/2-cytomegalovirus (CMV)-PsCatCh2.0-enhanced green fluorescent protein (EGFP) was injected into the vitreous cavity 1 mm below the corneoscleral limbus of mice in the rd1 experimental group, and the same dose of recombinant virus was injected 2 weeks later at temporal side 1 mm below the corneoscleral limbus. One year after virus injection, the light response of RGCs expressing PsCatCh2.0 was recorded by patch clamp technique; the expression of PsCatCh2.0 in the retina was evaluated by immunofluorescence staining; the transfection efficiency of recombinant virus was evaluated by the transfection efficiency of virus and the number of RGCs. Hematoxylin-eosin staining was performed to measure the inner retinal thickness. Western blotting was used to detect the protein expression of nuclear factor (NF)-κB p65 in retina; real-time quantitative polymerase chain reaction was used to detect the relative expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and Bax mRNA. Terminal deoxynucleotidyl transferase kit was used to observe the apoptosis of retinal cells in each group of mice. ResultsOne year after the intravitreal injection of recombinant virus, PsCatCh2.0-expressing RGCs can still generate 30 pA photocurrent. The virus PsCatCh2.0-EGFP was mainly transfected into RGCs, and partly transfected into amacrine cells, almost no transfection was seen in bipolar and horizontal cells. There were no significant differences in the number of RGCs and thickness of the inner retina between the rd1 experimental group and the rd1 control group (F=14.35, 0.05; P＞0.05), while the rd1 experimental group NF-κB p65 protein expression, TNF-α and IL-6 mRNA quantification were significantly lower than those of rd1 control group (F=4.61, 5.91, 5.78; P＜0.05). The number of red fluorescent apoptotic cells in the retina of mice in the rd1 experimental group was less than that in the rd1 control group, and the Bax mRNA expression was lower than that in the rd1 control group, and the difference was statistically significant (F=7.52, P＜0.01). ConclusionOne year after intravitreal injection of recombinant virus, the PsCatCh2.0 expressing RGCs can still generate photocurrent. Long term transfection and expression of PsCatCh2.0 has no obvious cytotoxic effect on RGCs, nor it increases the inflammatory effect of the retina of rd1 mice with retinal degeneration.
ObjectiveTo explore the light sensitivity and kinetic of the new optogenetics tools Channelrhodopsin-XXM2.0 (XXM2.0) and Channelrhodopsin-PsCatCh2.0 (PsCatCh2.0), and analyze whether they could be used to restore the visual function by optogenetics.MethodsMolecular biology techniques were used to link the gene fragments of XXM2.0 and PsCatCh2.0 to the vector pCIG(c)-msFoxn3 containing ampicillin resistant screening gene and reporter gene to form new plasmid pCIG(c)-msFoxn3-XXM2.0 and pCIG(c)-msFoxn3-PsCatCh2.0. The constructed plasmids were transfected into HEK 293T cells, and light responses were recorded in the whole cell mode with the HEKA patch clamp system. The photocurrent was recorded under three light intensity included 2.7×1016, 4.7×1015, and 6.4×1014 photons/(cm2·s). And then, XXM2.0 and PsCatCh2.0 were stimulated with 2.7×1016 photons/(cm2·s) and fully recovered. The opening and closing time constants were analyzed with Clampfit 10.6 software. At the same light intensity, photocurrents of XXM2.0 and PsCatCh2.0 were recorded by the light pulse stimulating of 2-32 Hz. The current attenuation was analyzed at long intervals of 4000 ms and short intervals of 200 ms after repeated stimulation. Comparisons between groups were performed by independent samples t test.ResultsRestriction endonuclease sites of EcoRⅠ and EcoRⅤ were successfully introduced at XXM2.0 and PsCatCh2.0 sequences. When the digestion was completed, they were ligated by T4 DNA ligase to construct new plasmids pCIG(c)-msFoxn3-XXM2.0 and pCIG (c)-msFoxn3-PsCatCh2.0, and then transfected on HEK 293T cells. The light intensity dependence was showed in XXM2.0 and PsCatCh2.0. The greater light intensity was accompanied by the greater photocurrent. Under the light intensity 6.4×1014 photons/(cm2·s) below the retinal safety threshold, large photocurrent was still generated in XXM2.0 and PsCatCh2.0 with 92.8±142.0 and 13.9±5.6 pA (t=1.24, 1.24; P=0.28, 0.29). The opening time constants of XXM2.0 and PsCatCh2.0 were 23.9±6.7 and 2.4±0.8 ms, and the closing time constants were 5803.0±568.2 and 219.9±25.6 ms. Compared with PsCatCh2.0, the opening and closing time constant of XXM2.0 were both larger than PsCatCh2.0. The differences were statistically significant (t=7.10, 31.60; P=0.00, 0.00). In terms of response frequency, XXM2.0 and PsCatCh2.0 could follow to 32 Hz high-frequency pulsed light stimulation, and all could respond to repeated light stimulation at a long (4000 ms) and a short time (200 ms) interval with the small current decay rate.ConclusionXXM2.0 and PsCatCh2.0 could be activated under light intensity with safety for the retina, and could respond to high frequency (at least 32 Hz) pulsed light stimuli with low current attenuation, which could meet the characteristics of opsins required to restore the visual function by optogenetics.
PURPOSE: To explore the pathogenesis of anisometropic and amblyopias. METHODS:To carry out on monocular and binocular atropinized cat models during the developmental period for anisometropia and ametropia ,and measure the cytosomal sectional area and some parameters of the dendric field from the dorsal lateral geniculate nuclei (dLGN)of adult cats by using Golgi-Cox staining. RESULIS:The changes of cytosomal sectional areas and parameters about dendric fields in the dLGN of experimental cats were as following:significant differences between cells of dLGN's A1 lamina by the monocular atropinized eyes and normal ones, binocular atropinized eyea and normal ones;no significant difference between tbat driven by the monoular and binocular atropinized eyes. CONCLUSIONS:There might be resemble pathogenesis between anisomelropic and ametropic amblyopias. （Chin J Ocul Fundus Dis,1996,12:153-156）
ObjectiveTo observe the expression of Rap1, guanosine triphosphate-Rap1 (GTP-Rap1), vascular endothelial growth factor (VEGF) and β-catenin in experimental choroidal neovascularization (CNV).MethodsForty-two brown Norwegian rats were randomly divided into a blank control group (7 rats) and a model group (35 rats). Both eyes were enrolled. The CNV model was established by holmium ion laser photocoagulation in the model group. At 3, 7, 14, 21, and 28 days after photocoagulation, fluorescein fundus angiography (FFA) and choroidal vascular smear were performed to observe the degree of fluorescein leakage and CNV area in rats; Western blot and real-time quantitative polymerase chain reaction (RT-PCR) were used to detect the expression of Rap1, GTP-Rap1, VEGF, β-catenin and mRNA in CNV.ResultsThe results of FFA examination showed that a large disc-shaped fluorescein leaked in the photo-condensation spot 14 days after photocoagulation. Laser confocal microscopy showed that compared with 7 days after photocoagulation, CNV area increased at 14, 21, 28 days after photocoagulation, and the difference were statistically significant (t=3.725, 5.532, 3.605;P＜0.05). Western blot showed that there was no significant difference in the relative expression of Rap1 protein in CNV at different time points after photocoagulation between the two groups (P=0.156). Compared with the blank control group, the relative expression of GTP-Rap1 protein was significantly decreased, the relative expression of VEGF and β-catenin protein were significantly increased in the model group (P=0.000). The results of RT-PCR showed that there was no significant difference in the relative expression of Rap1 mRNA at different time points after photocoagulation between the two groups (P=0.645), but there were significant difference in the relative expression of β-catenin mRNA (P=0.000). At 7, 14, 21 and 28 days after photocoagulation, there were significant difference in the relative expression of GTP-Rap1 and VEGF mRNA between the two groups (P=0.000).ConclusionsThe expression of GTP-Rap1 in experimental CNV is significantly lower than that in normal rats.
Objective To study the effects of long term application of cathartics on electromyography of rat colon, and to explore the role of interstitial cells of Cajal (ICC) in it. Methods Colonic slow waves of the rat was examined after 3-month feeding of phenolphthalein, and ICC in myenteric plexus was observed by ZIO method, and ultrastructure changes of nerves and ICC was observed. Results The frequency of slow waves of cathartic colon was reduced significantly(Ｐ＜0.05). The distribution of ICC in myenteric plexus was uneven, and the processes were mussily connected each other. Vacuolar degeneration of axon and ICC-like cells was revealed by electron microscope in myenteric plexus of cathartic colon. Conclusion Long term application of phenolphthalein could reduce the frequency of colonic slow waves, and the possible mechanism was degeneration of ICC and myenteric plexus nerves.
Objective To investigate the expression of cerb B2 and CathepsinD in gastric carcinoma and its correlation with the biological behavior of gastric carcinoma (GC). MethodsThe expression was studied by immunohistochemical technique. The expression of cerb B2 and CathepsinD were analyzed with their relation to histologic types, depth of invasion, growth pattern, lymph node metastasis and prognosis of gastric carcinoma. ResultsThirtynine of the 102 gastric carcinoma specimens (38.24%) were positive for cerb B2 and correlated with depth of invasion (P＜0.05) and lymph node metastasis (P＜0.05); eightythree of the 102 gastric carcinoma specimens (81.37%) were positive for CathepsinD and correlated with depth of invasion (P＜0.05), growth pattern (P＜0.05), lymph node metastasis (P＜0.05) and blood vessels cancer embolus (P＜0.05). Prognosis of patients with gastric carcinoma with positive expression of cerb B2 or CathepsinD was poor. The 5year survival rate was significantly lower in gastric carcinoma patients with positive expression of cerb B2 or CathepsinD. Conclusion cerb B2 and CathepsinD are highly related to growth, invasion, metastasis and prognosis of gastric carcinoma.
ObjectiveTo explore the current status of nursing researches about catheter-related infections in recent 5 years in China, and provide reference for further research.MethodsThe China National Knowledge Infrastructure database and Wangfang database were selected to search for literature about catheter-related infections published in recent 5 years in 10 nursing journals of Statistic Source, with the terms of " catheter-related bloodstream infection” or " ventilator-associated pneumonia” or " catheter-related urinary tract infection”. Statistical analysis was conducted.ResultsA total of 216 papers were included. The number of papers was not increased year by year. In terms of the object of study, the studies on ventilator-associated pneumonia were the most, accounting for 71.3%. Only 36.5% of the studies had accurate diagnostic criteria. In terms of the type of study, experimental studies were the most (109 articles). The content was concentrated on the best practice intervention studies (149 articles). The data collection methods gave priority to active surveillance/screening, including 113 articles.ConclusionsThe present focused attention is not enough to the researches about catheter-related infections in nursing field, and rigorous design is lacking in published studies. There are only a few nursing studies about catheter-related infections with high quality and high level of evidence. The quantity and quality of nursing researches about catheter-related infections still need to be improved. Nurses should pay more attention to the control and prevention of catheter-related infections, and should improve their research capacity.
ObjectiveTo investigate the incidence and trendency of healthcare-associated infections (HAIs) in a pediatric intensive care unit (ICU) of a hospital, identify the main objectives of infection control, and formulate corresponding preventive and control measures.MethodsA prospective targeted monitoring method was adopted to investigate HAIs in the pediatric ICU of a hospital from January 2013 to December 2018.ResultsFrom January 2013 to December 2018, the number of target ICU patients was 11 898, the number of patient-days was 55 159; 226 HAIs occurred, the HAI case rate was 1.90%, the incidence of HAI per 1 000 patient-days was 4.10‰, and the adjusted incidence of HAI per 1 000 patient-days was 1.21‰. The main infection site was respiratory tract [83 cases (36.7%)], with ventilator-associated pneumonia in 73 cases (32.3%); secondly, 69 patients (30.5%) had bloodstream infection, among which 48 (21.2%) had non-catheter-related bloodstream infection.ConclusionHospital targeted monitoring is helpful to grasp the situation and trend of HAIs, define the main target of infection control, and formulate corresponding preventive and control measures, which can effectively reduce the incidence of HAIs.
【Abstract】Objective To investigate the diagnostic and therapeutic value of selective angiography and embolization treatment in massive biliary hemorrhage in emergency. Methods Eleven cases of massive biliary bleeding were selectively angiographied and embolized with thread segment and gelatin sponge grain by Seldinger technique in emergency. Results Massive biliary bleeding in this series included bleeding after biliary surgery (7), post-laparoscopic cholecystectomy bleeding (1), bleeding from traumatic injury to liver (1), and rupture of huge hepatic cancer (2). There were 9 cases with abnormal angiographic findings including contrast medium leakage (9), tumor vessels and tumor tissue staining (2) and signs of pseudo-aneurysm formation (4). All the bleedings were stopped after emergency intervention but 2 cases with recurrent massive bleeding after 48 hours. Conclusion Emergency trans-catheter embolization treatment is an effective and safe method for biliary massive hemorrhage. It is also important to keep the biliary tract unobstructed.
Abstract： Objective To analyze clinical outcomes of interventional therapy for common congenital heart diseases （CHD）. Methods We retrospectively analyzed the clinical records of 57 patients with CHD who underwent catheter interventional therapy in People’s Hospital of Deyang City between March 2009 and January 2012. There were 31 male patients and 26 female patients with their mean age of 29.4±3.5 years（ranging from 1.5 to 75.0 years）. There were 21 patients with patent ductus arteriosus （PDA）, 12 patients with ventricular septal defect （VSD）, and 20 patients with atrial septal defect（ASD）；and 2 patients with VSD plus ASD, 1 patient with VSD plus PDA, and 1 patient with ASD plus PDA. Occluder and supplying system made in China were used in the interventional therapy. For the patients with ASD plus PDA, PDA was blocked before ASD；for the patients with VSD plus PDA, PDA was blocked before VSD；and for the patients with VSD plus ASD, VSD was blocked before ASD. Results The duration of interventional therapy ranged from 30 to 90 minutes. The success rate of operation was 98.3%（56/57）. Operation was given up in an old female patient with ASD and chronic obstructive pulmonary disease because of pulmonary hypertension. The hospital stay was 3 -7 days. All the patients were followed up at the outpatient department for 1 month to 2 years by color Doppler echocardiogram, chest X-ray and electrocardiograph, and no complication occurred during follow-up. Conclusion With complete understanding of surgical indications and strict compliance with procedures, catheter interventional therapy is safe, minimal invasive, and effective in CHD treatment.